Amperometric Detection of Apoptosis by using p-Methoxyaniline-Conjugated Substrate for Caspase-3

作者：Sun Sixiang; Inoue Kumi Y^*; Shiomoto Shusaku; Takano Shinichiro; Ino Kosuke; Shiku Hitoshi; Matsue Tomokazu^*

来源：ChemElectroChem, 2017, 4(4): 941-946.

DOI：10.1002/celc.201600700

• apoptosis
• biosensing
• caspase-3
• peptide substrate
• amperometric detection

This manuscript reports on a novel substrate for the amperometric detection of cellular apoptosis, Asp-Glu-Val-Asp-p-methoxyaniline (DEVD-pMA). This substrate showed electrochemical inertia from -0.2 to 0.7V versus Ag/AgCl in phosphate-buffered saline (PBS), but was cleaved by caspase-3, releasing pMA, which was detected with cyclic voltammetry and chronoamperometry. Compared with p-aminophenol (pAP), pMA showed lower toxicity to the human hepatocellular carcinoma cell line (HepG2) and higher stability in PBS. By using this substrate, we successfully detected caspase-3 in the range of 0.31-2.5x10(-2) units/mL and cellular apoptosis in HepG2 cells by using simple amperometry without cell lysis. This work will help to develop a simple method for the electrochemical monitoring of apoptosis, which can be applied in biological research and drug development.

• 出版日期2017-4